LOCUS V026691 9067 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026691 VERSION V026691 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /codon_start=1 /note="recognized by the 2H10 single-chain llama nanobody" /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /transl_table=1 /translation="KNEQELLELDKWASL" /label="gp41 peptide" misc_feature 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2843..3763 /label="Nmnat2(NM_175460)" /note="Nmnat2(NM_175460)" /gene="Nmnat2" CDS 3770..3835 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3855..4354 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4375..4974 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" misc_feature 4988..5576 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5783..6416 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6545..6561) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6569..6585 /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(6593..6623) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6638..6659 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6947..7535) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7706..8566) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(8567..8671) /gene="bla" /label="AmpR promoter" polyA_signal 8719..8853 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag 2821 ctcaagcttc gaattcgcca ccatgaccga gaccacaaag acccacgtta tcctgctggc 2881 ctgcggcagc ttcaatccca tcactaaagg gcacattcag atgttcgaga gagccaggga 2941 ttatctgcac aagactggaa gatttattgt gattggcggg attgtctctc cggtccatga 3001 ctcctacgga aaacagggcc ttgtgtcaag tcggcaccgt ctcatcatgt gtcagctggc 3061 tgtccagaat tccgactgga tcagggtgga cccatgggag tgctatcagg acacctggca 3121 gacaacctgc agtgtgttgg agcaccatcg agacctgatg aagagggtga ccggctgcat 3181 cctctccaac gtcaacacac cttccatgac acctgtgatc ggacagccac agcatgagaa 3241 cacccagccc atttaccaga acagcaatgt gcccaccaag cccactgcag ccaagatctt 3301 gggaaaggtg ggagaaagcc tcagccggat ctgctgcgtc cgcccaccag tggagcgctt 3361 cacttttgta gatgagaacg ccaacctggg cacagtgatg cggtatgagg agatcgagct 3421 gcgcatcttg ctgctgtgtg gtagtgacct gctggagtct ttctgcatcc caggactctg 3481 gaatgaggca gatatggaag tgattgttgg ggactttggg atcgtcgtgg tacccaggga 3541 tgcagcggac acagaccgga tcatgaatca ctcctccata ctccgcaagt acaaaaacaa 3601 catcatggtt gtgaaggatg atatcaacca tcccatgtct gtagtcagct ccaccaagag 3661 caggctggcc ctgcagcatg gggacggcca tgttgtggat tacctgtccc agccagtcat 3721 cgattacatc ctcaagagtc agctgtacat caacgcctcg ggcggatccg actacaaaga 3781 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtc 3841 tagataattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt 3901 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac 3961 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac 4021 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca 4081 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa 4141 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca 4201 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg 4261 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc 4321 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc 4381 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 4441 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 4501 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 4561 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 4621 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 4681 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 4741 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 4801 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 4861 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 4921 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaccgcgt 4981 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 5041 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 5101 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 5161 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 5221 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 5281 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 5341 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg 5401 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 5461 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 5521 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg 5581 aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca 5641 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca 5701 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac 5761 tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc 5821 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca 5881 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag 5941 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc 6001 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta 6061 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga 6121 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact 6181 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc 6241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 6301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 6361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag 6421 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag 6481 tgagaggcct tgacattgct agcgttttac cgtcgacctc tagctagagc ttggcgtaat 6541 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 6601 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 6661 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 6721 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 6781 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 6841 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 6901 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 6961 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 7021 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 7081 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 7141 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 7201 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 7261 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 7321 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 7381 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 7441 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 7501 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 7561 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 7621 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 7681 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 7741 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 7801 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 7861 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 7921 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 7981 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 8041 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 8101 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 8161 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 8221 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 8281 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 8341 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 8401 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 8461 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 8521 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 8581 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 8641 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 8701 tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat 8761 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 8821 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat 8881 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta 8941 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata 9001 tgtactacaa acttagtagt ttttaaagaa attgtatttg ttaaatatgt actacaaact 9061 tagtagt //